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bioRxiv · 10.1101/2023.02.15.528623

The thermostability of a VADEX-Pro based protein nanoparticle

Kan, M.-C.
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Abstract

We have adapted split GFP technology into the protein nanoparticle platform, Vaccine Delivery system X (VADEX), created in previous study. To evaluate the capability of this new platform, a model protein, maltose binding protein (MBP), was fused to the {beta}-strand 11 of sfGFP and co-expressed with VADEX-10 which was composed of LYRRLE peptide and N-terminal part up to {beta}-strand 10 of sfGFP. When these two fusion proteins were expressed in a cell, they were assembled into PNP spontaneously with a dynamic light scattering (DLS) particle size of 26 nm. This nanoparticle platform was renamed as VADEX-Pro for its capacity of expressing large protein on PNP. The thermostability of the assembled PNP was verified by both SDS-PAGE and DLS analysis following treatment. This PNP was stable at 25 {degrees}C and at temperatures as high as 40 {degrees}C for at least two months. Mutations that replaced cysteine residue of the LYRRLE peptide with serine or alanine destabilized and induced degradation of the VADEX-based PNP. The results in this study showed that the non-covalent complementation of split sfGFP became irreversible when reconstituted sfGFP was assembled in a VADEX-Pro PNP. This platform may be applied in developing thermostable vaccines.

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Kan, M.-C.. 2023-02-15. The thermostability of a VADEX-Pro based protein nanoparticle. https://doi.org/10.1101/2023.02.15.528623

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